rabbit anti rab11 d4f5  (Cell Signaling Technology Inc)

Journal: Cell reports

Article Title: Loss of the proton-activated chloride channel in neurons impairs AMPA receptor endocytosis and LTD via endosomal hyper-acidification

doi: 10.1016/j.celrep.2025.115302

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Article Snippet: Rabbit anti-Rab11 (D4F5) , Cell Signaling Technology , Cat. 5589, RRID: AB_10693925.

Techniques: Virus, Plasmid Preparation, Recombinant, shRNA, Control, Software

Journal: iScience

Article Title: A competition network connects Rab5 and Rab11 GTPases at the surface of endocytic structures

doi: 10.1016/j.isci.2025.112170

Figure Lengend Snippet: Measure of Rab5 and Rab11 functional interactions using FRET imaging and correlation analysis (A) Schematic representation of the imaging procedure employed to measure Rab5 and Rab11 functional interaction using Rab5 and Rab11 fluorescent probes and activity sensors. The square box represents the magnification of schematic spherical endosomes with Rab5 and Rab11 fluorescent tagged proteins. (B) Representative localization images of both Rab5 FRET biosensor (green) and mCherry-Rab11 (magenta) in the same single Cos-7 cell. Scale bars for whole cell (left panel) and the enlarged spherical endosome (right panel) are shown. (C) Quantification of both Rab5-positive (top panel) and Rab11-positive (bottom panel) structures motility in Cos-7 cells (mean square displacement, MSD). n = 198 Rab5-positive structures ans n = 322 Rab11-positive structures. Black arrow indicates static structures used for subsequent analysis. (D) Representative correlation profiles computed for different combinations of both Rab5 and Rab11 fluorescence intensity (I) and FRET efficiency (E) values measured at the surface of a single spherical endosomes. Dashed and solid lines represent control and Rab11FIP5 expressing cells, respectively (4 trajectories per condition are displayed). n = 12 endosomes for cells co-transfected with Rab5 FRET biosensor, mCherry-Rab11 and either control or Rab11FIP5 plasmids. n = 13 endosomes for cells co-transfected with Rab11 FRET biosensor, mCherry-Rab5 and control plasmid; n = 11 endosomes for cells co-transfected with Rab11 FRET biosensor, mCherry-Rab5 and Rab11FIP5. (E) Representative correlation profiles computed for different combinations of both Rab5 and Rab11 fluorescence intensity (I) and FRET efficiency (E) values measured at the surface of a single spherical endosomes. Dashed and solid lines represent control and Zfyve26 expressing cells, respectively (4 trajectories per condition are displayed). n = 16 endosomes for cells co-transfected with Rab5 FRET biosensor, mCherry-Rab11 and either control or Zfyve26 plasmids. n = 8 endosomes for cells co-transfected with Rab11 FRET biosensor, mCherry-Rab5 and control plasmid. n = 9 endosomes for cells co-transfected with Rab11 FRET biosensor, mCherry-Rab5 and Zfyve26 plasmids.

Article Snippet: Rab11 (D4F5) , Cell Signaling Technology , Cat# 5589; RRID: AB_10692655.

Techniques: Functional Assay, Imaging, Activity Assay, Fluorescence, Control, Expressing, Transfection, Plasmid Preparation

Journal: iScience

Article Title: A competition network connects Rab5 and Rab11 GTPases at the surface of endocytic structures

doi: 10.1016/j.isci.2025.112170

Figure Lengend Snippet: Quantification of Pearson correlations of Rab5 and Rab11 GTPases at the surface of a single endosome

Article Snippet: Rab11 (D4F5) , Cell Signaling Technology , Cat# 5589; RRID: AB_10692655.

Techniques: Control

Journal: iScience

Article Title: A competition network connects Rab5 and Rab11 GTPases at the surface of endocytic structures

doi: 10.1016/j.isci.2025.112170

Figure Lengend Snippet: Functional impact of Zfyve26 on endogenous Rab5 and Rab11 (A) Representative images of spherical endosomes stained for endogenous Rab5 (green), endogenous Rab11 (magenta) and GFP-Zfyve26 (blue). Scale bars for merged image (left panel) and single channels (right panels) are displayed. (B) Number of Rab5 (left panel) and Rab11 (right panel) structures per single cell. Boxplots indicate median (middle line), 25th, 75th percentile (box) and 5th and 95th percentile (whiskers) as well as outliers (single points). ∗ p value <0.05 (Kolmogorov-Smirnov test), n = 102 cells in GFP expressing condition (control), n = 97 cells in GFP-Zfyve26 expressing condition (Zfyve26). (C) Quantification of Rab5 (left panel) and Rab11 mean intensity (right panel) per single spherical endosome. Error bars indicate mean ± S.E.M. ∗∗ p value <0.01, ∗∗∗∗ p value <0.0001 (Student’s t test), n = 26 spherical endosomes in GFP expressing condition (control) and n = 22 spherical endosomes GFP-Zfyve26 expressing condition (Zfyve26). (D) Quantification of Rab5 (left panel) and Rab11 structures area (right panel) per single spherical endosomes. Error bars indicate mean ± S.E.M. ns = not significant, n = 26 spherical endosomes in GFP expressing condition (Control) and n = 22 spherical endosomes in GFP-Zfyve26 expressing condition (Zfyve26). (E) Quantification of Rab5-Rab11 separation per single spherical endosome. Rab5-Rab11 separation is computed as 1 minus Rab5-Rab11 Mander’s coefficient. Error bars indicate mean ± S.E.M. ∗ p value <0.05 (Student’s t test), n = 26 spherical endosomes in GFP expressing condition (Control) and n = 22 spherical endosomes in GFP-Zfyve26 expressing condition (Zfyve26). (F) Quantification of endogenous Rab5-GTP content in cells upon Zfyve26 expression by using Rab5 activation pull-down assay. Representative western blot images (left panel) and quantification (right panel). FC indicate fold-change over control. Error bars indicate mean ± S.E.M. ∗ p value <0.05 (Student’s t test), n = 3 independent experiments. (G) Quantification of transfected Rab5-GTP content in cells upon expression of Rab5 variants and/or Zfyve26 by using Rab5 activation pull-down assay. Representative western blot images (left panel) and quantification (right panels). FC indicate fold-change over control. Error bars indicate mean ± S.E.M. ∗ p value <0.05 (Student’s t test), n = 4 independent experiments. (H) Quantification of endogenous Rab11-GTP content in cells upon Zfyve26 expression by using Rab11 activation pull-down assay. Representative western blot images (left panel) and quantification (right panel). FC indicate fold-change over control. Error bars indicate mean ± S.E.M. ∗ p value <0.05 (Student’s t test), n = 3 independent experiments. (I) Quantification of transfected Rab11-GTP content in cells upon expression of Rab11 variants and/or Zfyve26 by using Rab11 activation pull-down assay. Representative western blot images (left panel) and quantification (right panels). FC indicates fold-change over control. Error bars indicate mean ± S.E.M. ns = not significant, ∗ p value <0.05 (Student’s t test), n = 4 independent experiments. (J) Quantification of transferrin recycling using Alexa Fluor 647-conjugated human transferrin upon GFP (control) or GFP-Zfyve26 overexpression. Representative images (left panels) and quantification (right panel) (scale bar, 10 μm). Data represents mean ± S.E.M., ∗∗∗ p value <0.001 (Student’s t test), n = 3 independent experiments.

Article Snippet: Rab11 (D4F5) , Cell Signaling Technology , Cat# 5589; RRID: AB_10692655.

Techniques: Functional Assay, Staining, Expressing, Control, Activation Assay, Pull Down Assay, Western Blot, Transfection, Over Expression

Journal: iScience

Article Title: A competition network connects Rab5 and Rab11 GTPases at the surface of endocytic structures

doi: 10.1016/j.isci.2025.112170

Figure Lengend Snippet: Quantification of both abundance and activity of Rab5 and Rab11 GTPases at the surface of a single endosome

Article Snippet: Rab11 (D4F5) , Cell Signaling Technology , Cat# 5589; RRID: AB_10692655.

Techniques: Activity Assay, Control

Journal: iScience

Article Title: A competition network connects Rab5 and Rab11 GTPases at the surface of endocytic structures

doi: 10.1016/j.isci.2025.112170

Figure Lengend Snippet: Zfyve26 localization on endosomes relies on phosphatidylinositol 3-phosphate (A) Quantification of fluorescence recovery after photo bleaching (FRAP) on GFP-Zfyve26-positive spherical endosomes. Representative image of a cell expressing GFP-Zfyve26 (left). Quantification and representative time course images of a photobleaching experiments (right panels). Gray area represents standard error of mean (SEM) of data. n = 20 GFP-Zfyve26-positive spherical endosomes. (B) Representative western blot of Zfyve26 binding to recombinant GDP- or GTPγS-bound Rab5 by pull-down assay (left panel), n = 3 independent experiments. Representative western blot of Zfyve26 binding to recombinant GDP- or GTPγS-bound Rab11 by pull-down assay (right panel), n = 3 independent experiments. (C) Quantification of the number of GFP-Zfyve26 positive structures per single cell upon treatment with VPS34-IN1, a PIK3C3 inhibitor. Representative images of GFP-Zfyve26 positive structures in control (vehicle, top left panel) or treated (VPS34-IN1, bottom left panel) conditions and quantification (right panel). Boxplots indicate median (middle line), 25th, 75th percentile (box) and 5th and 95th percentile (whiskers). ∗∗∗∗ p value <0.0001 (Kolmogorov-Smirnov test), n = 61 cells in control condition, n = 67 cells in treated cells. (D) Quantification of the number of GFP-Zfyve26 structures in cells co-expressing GFP-Zfyve26/mCherry (control), GFP-Zfyve26/mCherry-Rab5(S34N) (Rab5(S34N)), GFP-Zfyve26/mCherry-Rab5(Q79L) (Rab5(Q79L)),treated with either DMSO (vehicle) or VPS34-IN1 inhibitor (VPS34-IN1). Boxplots indicate median (middle line), 25th, 75th percentile (box) and 5th and 95th percentile (whiskers) as well as outliers (single points). ∗∗∗∗ p value <0.0001 (Kolmogorov-Smirnov test), n = 50 cells in control condition (vehicle), n = 48 cells in Rab5(S34N) condition (vehicle), n = 48 cells in Rab5(Q79L) condition (vehicle), n = 50 cells in control condition (VPS34-IN1), n = 55 cells in Rab5(S34N) condition (VPS34-IN1), n = 50 cells in Rab5(Q79L) condition (VPS34-IN1).

Article Snippet: Rab11 (D4F5) , Cell Signaling Technology , Cat# 5589; RRID: AB_10692655.

Techniques: Fluorescence, Expressing, Western Blot, Binding Assay, Recombinant, Pull Down Assay, Control

Journal: iScience

Article Title: A competition network connects Rab5 and Rab11 GTPases at the surface of endocytic structures

doi: 10.1016/j.isci.2025.112170

Figure Lengend Snippet: Correlations analysis captures Zfyve26 saturation at the endocytic surface by using loop index (A) Representative magnification of the Rab5 Loop IDX at the surface of a single endocytic organelle resulting from kymograph data in both control (top panels) and Zfyve26 expressing cells (bottom panels). Black-to-green, blue-to-yellow and blue-to-orange gradients represent Rab5 abundance, Rab5 FRET efficiency and Rab5 Loop IDX, respectively. (B) Representative magnification of the Rab11 Loop IDX at the surface of a single endocytic organelle resulting from kymograph data in both control (top panels) and Zfyve26 expressing cells (bottom panels). Black-to-magenta, blue-to-yellow and blue-to-orange gradients represent Rab11 abundance, Rab11 FRET efficiency and Rab11 Loop IDX, respectively. (C) Scatterplot of both Rab5 activity (i.e., Rab5 ( E )) and Rab5 Loop IDX measured for both control and Zfyve26 expressing cells transfected with Rab5 activity sensor. White-to-green gradient represents data density for control cells. White-to-dark green gradient represents data density for Zfyve26 expressing cells. Lines represent the correlation (Saturation, Sat.) values ( n = 10800 subsampled values). (D) Scatterplot of subsampled RabX activity and RabX Loop IDX at different effector level (0, 50, 200, and 400) using model III. Data magnification for high (top left, 400) and low (bottom left, 50) effector values are displayed. Dots of different green shades represent values obtained at different effector level. (n simulations = 5 × 10, subsampled in batches of n = 50). (E) Scatterplot of both Rab11 activity (i.e., Rab11 ( E )) and Rab11 Loop IDX measured for both control and Zfyve26 expressing cells transfected with Rab11 activity sensor. White-to-magenta gradient represents data density for control cells. White-to-dark magenta gradient represents data density for Zfyve26 expressing cells. Lines represent the correlation (Saturation, Sat.) values ( n = 10800 subsampled values). (F) Scatterplot of subsampled RabY activity and RabY Loop IDX at different effector level (0, 50, 200, and 400) using model III. Data magnification for high (top left, 400) and low (bottom left, 50) effector values are displayed. Dots of different magenta shades represent values obtained at different effector level. (n simulations = 5 × 10, subsampled in batches of n = 50).

Article Snippet: Rab11 (D4F5) , Cell Signaling Technology , Cat# 5589; RRID: AB_10692655.

Techniques: Control, Expressing, Activity Assay, Transfection

Journal: iScience

Article Title: A competition network connects Rab5 and Rab11 GTPases at the surface of endocytic structures

doi: 10.1016/j.isci.2025.112170

Figure Lengend Snippet:

Article Snippet: Rab11 (D4F5) , Cell Signaling Technology , Cat# 5589; RRID: AB_10692655.

Techniques: Virus, Recombinant, Imaging, Software, Microscopy

Journal: Communications Biology

Article Title: Involvement of sphingosine 1-phosphate signaling in insulin-like growth factor-II/mannose 6-phosphate receptor trafficking from endosome to the trans -Golgi network

doi: 10.1038/s42003-024-06828-9

Figure Lengend Snippet: A HeLa cells were incubated with 5 µM HACPT, 5 µM W146, 5 µM VPC23019 or DMSO as a vehicle in the presence of cycloheximide containing standard medium for 3 h and fixed for immunofluorescence study with anti-IGF-II/M6P receptor and anti-golgin-97 antibodies. Cells were analyzed by confocal microscopy. Cell boundaries are indicated as dotted lines. B The 30 cell images were subjected for Pearson’s coefficient analysis. Results were expressed as the mean ± s.e.m. (** p < 0.01 versus vehicle treatment; Welch’s t -test) on scatter-dot plots. C HeLa cells were transfected with SphK1-, SphK2-, S1P 1 R-, S1P 3 R- or VPS35-siRNA and fixed, stained and analyzed as in ( B ). Microscopic data were shown in Supplementary Fig. . Bars represent the mean ± s.e.m. (* p < 0.05; ** p < 0.01 versus control siRNA; Welch’s t -test). D HeLa cells were incubated with 5 µM VPC23019 in the presence of cycloheximide containing standard medium for 3 h and fixed for immunofluorescence study with anti-IGF-II/M6P receptor, anti-Golgin97 and anti-Rab5, anti-Rab7 or anti-Rab11 antibodies. Note that some IGF-II/M6P receptor-positive endosomes outside TGN were co-localized clearly with Rab5 (see arrows), but only slightly with Rab11 and hardly with Rab7. The cell boundaries are indicated as dotted lines. Scale bars, 10 µm.

Article Snippet: Anti-CD8a antibody (Clone RPA-T8) was from BioLegend; rabbit monoclonal anti-CD8a antibody (clone 1B19) was from Merck; rabbit anti-golgin-97 antibody from Proteintech; mouse anti-golgin97 antibody (clone CDF4) from Molecular Probes; anti-Rab5 (clone C8B1), anti-Rab7 (clone D95F2) and anti-Rab11 (clone D4F5) antibodies from Cell Signaling Technologies; anti-DDDDK-tag antibody and anti-α-tubulin antibody from Medical & biological laboratories; anti-VPS35 (clone B-5), anti-CK2α (clone E-7) and anti-SphK1 (clone G-11) from Santa Cruz; rabbit anti-M6PR antibody (clone EPR6599) from Abcam; mouse anti-M6PR antibody (clone 2G11) from Fitzgerald; anti-β-actin antibody from WAKO.

Techniques: Incubation, Immunofluorescence, Confocal Microscopy, Transfection, Staining, Control

Journal: Neuron

Article Title: Rapid iPSC inclusionopathy models shed light on formation, consequence, and molecular subtype of α-synuclein inclusions

doi: 10.1016/j.neuron.2024.06.002

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-Rab11, clone D4F5 , Cell Signaling , Cat. # 5589; RRID:AB_10693925.

Techniques: Transduction, CRISPR, Sequencing, Software